title: Exocrine cell-derived microparticles in response to lipopolysaccharide promote endocrine dysfunction in cystic fibrosis.
abstract: BACKGROUND
  Diabetes in cystic fibrosis (CF) is a result of exocrine pancreas alteration followed by endocrine dysfunction at a later stage. Microparticles (MPs) are plasma membrane fragments shed from stimulated or damaged cells that act as cellular effectors. Our aim was to identify a new form of interaction between exocrine and endocrine pancreatic cells mediated by exocrine MPs, in the context of recurrent infection in CF.
  
  
  METHODS
  MPs from either human exocrine CFTRΔF508-mutated (CFPAC-1) cells or exocrine normal pancreatic (PANC-1) cells were collected after treatment by LPS from Pseudomonas aeruginosa and applied to rat endocrine normal insulin-secreting RIN-m5F cells. MP membrane integration in target cells was established by confocal microscopy and flow cytometry using PKH26 lipid probe. Apoptosis, lysosomal activity, insulin secretion were measured after 18 h. MP-mediated NF-κB activation was measured in HEK-Blue reporter cells by SEAP reporter gene system and in RIN-m5F cells by Western blot. In endocrine normal cells, CFTR inhibition was achieved using Inhibitor-172.
  
  
  RESULTS
  Compared to PANC-1, MPs from CFPAC-1 significantly reduced insulin secretion and lysosomal activity in RIN-m5F. MPs induced NF-κB activation by increasing the level of IκB phosphorylation. Moreover, the inhibition of NF-κB activation using specific inhibitors was associated with a restored insulin secretion. Interestingly, CFTR inhibition in normal RIN-m5F cells promoted apoptosis and decreased insulin secretion.
  
  
  CONCLUSIONS
  During recurrent infections associated with CF, exocrine MPs may contribute to endocrine cell dysfunction via NF-κB pathways. Membrane CFTR dysfunction is associated with decreased insulin secretion.
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, Strasbourg, France; Department of Parasitology and Parasitic Diseases and Animal Biology, Faculty of Veterinary Medicine, University of Agronomical Sciences and Veterinary Medicine, 105 spl. Independentei, sector 5, 050097 Bucharest, Romania.
authors.forename: Andrei Alexandru
authors.initials: AA
authors.lastname: Constantinescu
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, Strasbourg, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Gleizes
authors.affiliation: UMR7213 CNRS, Laboratory of Biophotonics and Pharmacology, 74 route du Rhin, F-67401 Illkirch, France.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Alhosin
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, Strasbourg, France.
authors.forename: Elhassan
authors.initials: E
authors.lastname: Yala
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, Strasbourg, France.
authors.forename: Fatiha
authors.initials: F
authors.lastname: Zobairi
authors.affiliation: Department of Pneumology, University Hospital, 1 place de L'hôpital, CHU de Strasbourg, BP426, 67091 Strasbourg Cedex, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Leclercq
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Bucharest, 91-95 spl. Independentei, sector 5, 050095 Bucharest, Romania.
authors.forename: Gheorghe
authors.initials: G
authors.lastname: Stoian
authors.affiliation: Department of Parasitology and Parasitic Diseases and Animal Biology, Faculty of Veterinary Medicine, University of Agronomical Sciences and Veterinary Medicine, 105 spl. Independentei, sector 5, 050097 Bucharest, Romania.
authors.forename: Ioan Liviu
authors.initials: IL
authors.lastname: Mitrea
authors.affiliation: EA7290 Early Bacterial Virulence, Faculty of Medicine, University of Strasbourg, 3 rue Koeberlé, F-67000 Strasbourg, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Prévost
authors.affiliation: UMR7213 CNRS, Laboratory of Biophotonics and Pharmacology, 74 route du Rhin, F-67401 Illkirch, France; Faculty of Pharmacy, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Toti
authors.affiliation: EA7293, Vascular and Tissular Stress in Transplantation, Federation of Translational Medicine of Strasbourg, Faculty of Medicine, University of Strasbourg, 74 route du Rhin, F-67401 Illkirch, Strasbourg, France; Department of Diabetology, University Hospital, 1 place de l'Hôpital, CHU de Strasbourg, BP421, 67091 Strasbourg Cedex, France. Electronic address: kesslerl@unistra.fr.
authors.forename: Laurence
authors.initials: L
authors.lastname: Kessler
mesh_terms: D000818:Animals; D002450:Cell Communication; D002460:Cell Line; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000150:complications / Q000378:metabolism / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D003920:Diabetes Mellitus / Q000209:etiology / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D046790:Pancreas, Exocrine / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000150:complications / Q000378:metabolism / Q000503:physiopathology; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D051381:Rats; D012008:Recurrence; D015398:Signal Transduction
chemical_list: C579654:CFTR protein, rat; D007328:Insulin; D008070:Lipopolysaccharides; D016328:NF-kappa B; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: Cystic fibrosis-related diabetes; Insulin secretion; Membrane microparticles; NF-kB